Single domain antibodies against enteric pathogen virulence factors are active as curli fiber fusions on probiotic E. coli Nissle 1917.

Autor: Gelfat I; John A. Paulson School of Engineering and Applied Sciences, Harvard University, Allston, Massachusetts, United States of America.; Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts, United States of America., Aqeel Y; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Tremblay JM; Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America., Jaskiewicz JJ; Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America., Shrestha A; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Lee JN; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Hu S; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Qian X; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Magoun L; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Sheoran A; Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America., Bedenice D; Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America., Giem C; Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts, United States of America., Manjula-Basavanna A; Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts, United States of America., Pulsifer AR; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Tu HX; Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts, United States of America., Li X; Department of Mechanical and Industrial Engineering, Northeastern University, Boston, Massachusetts, United States of America., Minus ML; Department of Mechanical and Industrial Engineering, Northeastern University, Boston, Massachusetts, United States of America., Osburne MS; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America., Tzipori S; Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America., Shoemaker CB; Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University, North Grafton, Massachusetts, United States of America., Leong JM; Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts, United States of America.; Stuart B. Levy Center for Integrated Management of Antimicrobial Resistance, Tufts University, Medford, Massachusetts, United States of America., Joshi NS; Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts, United States of America.
Jazyk: angličtina
Zdroj: PLoS pathogens [PLoS Pathog] 2022 Sep 15; Vol. 18 (9), pp. e1010713. Date of Electronic Publication: 2022 Sep 15 (Print Publication: 2022).
DOI: 10.1371/journal.ppat.1010713
Abstrakt: Enteric microbial pathogens, including Escherichia coli, Shigella and Cryptosporidium species, take a particularly heavy toll in low-income countries and are highly associated with infant mortality. We describe here a means to display anti-infective agents on the surface of a probiotic bacterium. Because of their stability and versatility, VHHs, the variable domains of camelid heavy-chain-only antibodies, have potential as components of novel agents to treat or prevent enteric infectious disease. We isolated and characterized VHHs targeting several enteropathogenic E. coli (EPEC) virulence factors: flagellin (Fla), which is required for bacterial motility and promotes colonization; both intimin and the translocated intimin receptor (Tir), which together play key roles in attachment to enterocytes; and E. coli secreted protein A (EspA), an essential component of the type III secretion system (T3SS) that is required for virulence. Several VHHs that recognize Fla, intimin, or Tir blocked function in vitro. The probiotic strain E. coli Nissle 1917 (EcN) produces on the bacterial surface curli fibers, which are the major proteinaceous component of E. coli biofilms. A subset of Fla-, intimin-, or Tir-binding VHHs, as well as VHHs that recognize either a T3SS of another important bacterial pathogen (Shigella flexneri), a soluble bacterial toxin (Shiga toxin or Clostridioides difficile toxin TcdA), or a major surface antigen of an important eukaryotic pathogen (Cryptosporidium parvum) were fused to CsgA, the major curli fiber subunit. Scanning electron micrographs indicated CsgA-VHH fusions were assembled into curli fibers on the EcN surface, and Congo Red binding indicated that these recombinant curli fibers were produced at high levels. Ectopic production of these VHHs conferred on EcN the cognate binding activity and, in the case of anti-Shiga toxin, was neutralizing. Taken together, these results demonstrate the potential of the curli-based pathogen sequestration strategy described herein and contribute to the development of novel VHH-based gut therapeutics.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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